Taze Kaşar Peynirinden İzole Edilen Lactobacillus brevis'ten Fitaz Enziminin Kısmi Saflaştırılması ve Biyokimyasal Karakterizasyonu

Öz

Son 20 yılda fitaz enzimi çevre koruma, beslenme ve biyoteknoloji alanlarında bilim insanlarının dikkatini çekmiştir. Fosfataz enzim grubunda yer alan miyo-inositol heksafosfat fosfohidrolaz (fitaz), fitatın daha az fosforile edilmiş inorganik fosfatlara ve fitata hidrolizini katalizler. Mikrobiyal kaynaklı fitazlar biyoteknolojik uygulamalarda (kâğıt endüstrisinde, yem sanayinde, gıda endüstrisinde ve toprak iyileştirmede) yaygın bir şekilde kullanılmaktadır. Mevcut çalışmada, taze kaşar peynirinden izole edilen Lactobacillus brevis NM-34 suşundan fitaz enzimi kısmi olarak saflaştırıldı ve enzimin optimum aktivite gösterdiği pH ve sıcaklık değerleri belirlendi. L. brevis NM-34 amonyum sülfat çöktürmesi sonucunda 243.80 U/mL’lik bir fitaz aktivitesi sergiledi. En yüksek fitaz aktivitesi görülen amonyum sülfat aralığında (%40-60) protein konsantrasyonu 0.989 mg/mL olarak ölçüldü. Fitaz enziminin Km ve Vmax değerleri sırasıyla 0.0146 mM ve 1.6 µmol/min olarak belirlendi. Kısmi olarak saflaştırılan fitazın optimum aktivite gösterdiği pH ve sıcaklık değerleri sırasıyla pH 5.0 ve 50 °C olarak bulundu. Araştırmamızdan elde edilen bulgulara dayanarak, bu bakteriden saflaştırılan enzimin endüstriyel uygulamalarda kullanılmasını uygun kılan kendine özgü niteliklere sahip olduğu tespit edilmiştir.

Anahtar Kelimeler

• Lactobacillus brevis
• Fitaz
• Enzim saflaştırması

Purification and Biochemical Characterization of Phytase Enzyme from Lactobacillus brevis Isolated from Fresh Kashar Cheese

Abstract

In the last 20 years, phytase enzyme has attracted the attention of scientists in the fields of environmental protection, nutrition and biotechnology. Myo-inositol hexaphosphate phosphohydrolase (phytase), which is a type of phosphatase enzyme, catalyzes the hydrolysis of phytate into less phosphorylated inorganic phosphates and phytate. Phytases of microbial origin are widely used in biotechnological applications (paper industry, feed industry, food industry and soil improvement). In the present study, phytase enzyme was partially purified from Lactobacillus brevis NM-34 strain isolated from fresh kashar cheese and the pH and temperature values at which the enzyme showed optimum activity were determined. L. brevis NM-34 showed a phytase activity of 243.80 U/mL as a result of ammonium sulfate precipitation. In the ammonium sulfate range (40-60%), where the highest phytase activity was observed, the protein concentration was measured as 0.989 mg/mL. Km and Vmax values of phytase enzyme were determined as 0.0146 mM and 1.6 µmol/min, respectively. The pH and temperature values at which the partially purified phytase showed optimum activity were found to be pH 5 and 50 °C, respectively. Based on the findings from our research, the enzyme purified from this bacterium was found to have unique properties that make it suitable for use in industrial applications.

Keywords

• Lactobacillus brevis
• Phytase
• Enzyme purification

Kaynakça

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